LOCUS       sequence-472118-        3194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BEVA Golden Gate cloning vector; BEVA2.0 Narrow host range plasmid 
            with I-SceI counterselectable site. L1-P15A-Gm-ISceI-ELT4; GG 
            assembly from pOGG004, pOGG009, pNDGG006, pGQ0023, pOGG014. GmR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3194)
  AUTHORS   Geddes BA, Williamson R, Schumacher J, Ardi A, Levin G, Cervenka E, 
            Huang R, diCenzo G
  TITLE     BEVA2.0: modular assembly of golden gate-compatible vectors with 
            expanded utility for genetic engineering.
  JOURNAL   Can J Microbiol. 2025 Jan 1;71:1-13. doi: 10.1139/cjm-2024-0246.
  PUBMED    39919271
REFERENCE   2  (bases 1 to 3194)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Can J 
            Microbiol."; date: "2025-01-1"; volume: "71:1-13. doi"; pages: " 
            10.1139/cjm-2024-0246"
COMMENT     Sequence Label: pQGG004
FEATURES             Location/Qualifiers
     source          1..3194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(68..154)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    242..263
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(278..283,284..301,302..308)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    316..332
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     321..343
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     340..356
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     340..356
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             352..675
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    369..425
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(426..443)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(426..442)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(435..457)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(651..670)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      780..874
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        join(883..888,889..905,906..911)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1100..1633
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     oriT            1827..1936
                     /note="incP origin of transfer"
     rep_origin      2319..2864
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     2765..2784
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2981..2998
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ttactgacca ttccaaggta gggataacag ggtaatttgg aaccgtagtc cagatctagg
       61 gcggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
      121 cccagtcttt cgactgagcc tttcgtttta tttgatgcct gccgaattcg gatccggagt
      181 gagaccgcag ctggcacgac aggtttgccg actggaaagc gggcagtgag cgcaacgcaa
      241 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
      301 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
      361 attacgccaa gcttgcatgc ctgcaggtcg actctagagg atccccgggt accgagctcg
      421 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
      481 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
      541 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
      601 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
      661 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
      721 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgacggtctc acgctcttgg
      781 actcctgttg atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct
      841 cggttgccgc cgggcgtttt ttattggtga gaatccaggc aattgacata agcctgttcg
      901 gttcgtaaac tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc
      961 gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt
     1021 gtacagccta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg
     1081 atgttatgga gcagcaacga tgttacgcag cagcaacgat gttacgcagc agggcagtcg
     1141 ccctaaaaca aagttaggtg gctcaagtat gggcatcatt cgcacatgta ggctcggccc
     1201 tgaccaagtc aaatccatgc gggctgctct tgatcttttc ggtcgtgagt tcggcgacgt
     1261 agccacctac tcccaacatc agccggactc cgattacctc gggaacttgc tccgtagtaa
     1321 gacattcatc gcgcttgctg ccttcgacca agaagcggtt gttggcgctc tcgcggctta
     1381 cgttctgccc aagtttgagc agccgcgtag tgagatctat atctatgatc tcgcagtctc
     1441 cggagagcac cggaggcagg gcattgccac cgcgctcatc aatctcctca agcatgaggc
     1501 caacgcgctt ggtgcttatg tgatctacgt gcaagcagat tacggtgacg atcccgcagt
     1561 ggctctctat acaaagttgg gcatacggga agaagtgatg cactttgata tcgacccaag
     1621 taccgccacc taacaattcg ttcaagccga gatcggcttc ccggccgcgg agttgttcgg
     1681 taaattgact acttttccgc tgcataaccc tgcttcgggg tcattatagc gattttttcg
     1741 gtatatccat cctttttcgc acgatataca ggattttgcc aaagggttcg tgtagacttt
     1801 ccttggtgta tccaacggcg tcagccgggc aggataggtg aagtaggccc acccgcgagc
     1861 gggtgttcct tcttcactgt cccttattcg cacctggcgg tgctcaacgg gaatcctgct
     1921 ctgcgaggct ggccggctac cgccggcgta acagatgagg gcaagcggat ggctgatgaa
     1981 accaagccaa ccaggaaggg cagcccacct atcaaggtgt actgccttcc agacgaacga
     2041 agagcgattg aggaaaaggc ggcggcggcc ggcatgagcc tgtcggccta cctgctggcc
     2101 gtcggccagg gctacaaaat cacgggcgtc gtggactatg agcacgtccg cgagctggcc
     2161 cgcatcaatg gcgacctggg ccgcctgggc ggcctgctga aactctggct caccgacgac
     2221 ccgcgcacgg cgcggttcgg tgatgccacg atcctcgccc tgctggcgaa gatcgactct
     2281 agctagaaat attttatctg attaataaga tgatcttctt gagatcgttt tggtctgcgc
     2341 gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc
     2401 tgagctacca actctttgaa ccgaggtaac tggcttggag gagcgcagtc accaaaactt
     2461 gtcctttcag tttagcctta accggcgcat gacttcaaga ctaactcctc taaatcaatt
     2521 accagtggct gctgccagtg gtgcttttgc atgtctttcc gggttggact caagacgata
     2581 gttaccggat aaggcgcagc ggtcggactg aacggggggt tcgtgcatac agtccagctt
     2641 ggagcgaact gcctacccgg aactgagtgt caggcgtgga atgagacaaa cgcggccata
     2701 acagcggaat gacaccggta aaccgaaagg caggaacagg agagcgcacg agggagccgc
     2761 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc
     2821 gtcagatttc gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg
     2881 ccctctcact tccctgttaa gtatcttcct ggcatcttcc aggaaatctc cgccccgttc
     2941 gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga
     3001 agcggaatat atcctgtatc acatattctg ctgacgcacc ggtgcagcct tttttctcct
     3061 gccacatgaa gcacttcact gacaccctca tcagtgccaa catagtaagc cagtatacac
     3121 tccgctagcg ctgatgtccg gcggtgcttt tgccgttacg caccaccccg tcagtagctg
     3181 aacaggaggg acag
//
